Targeting the Blind Spot of Polycationic Nanocarrier-Based siRNA Delivery
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- 作者：Mengyao Zheng ; Giovanni M. Pavan ; Manuel Neeb ; Andreas K. Schaper ; Andrea Danani ; Gerhard Klebe ; Olivia M. Merkel ; Thomas Kissel
- 刊名：ACS Nano
- 出版年：2012
- 出版时间：November 27, 2012
- 年：2012
- 卷：6
- 期：11
- 页码：9447-9454
- 全文大小：512K
- 年卷期：v.6,no.11(November 27, 2012)
- ISSN：1936-086X
文摘
Polycationic nanocarriers attract increasing attention to the field of siRNA delivery. We investigated the self-assembly of siRNA vs pDNA with polycations, which are broadly used for nonviral gene and siRNA delivery. Although polyethyleneimine (PEI) was routinely adopted as siRNA carrier based on its efficacy in delivering pDNA, it has not been investigated yet why PEI efficiently delivers pDNA to cells but is controversially discussed in terms of efficacy for siRNA delivery. We are the first to investigate the self-assembly of PEI/siRNA vs PEI/pDNA and the steps of complexation and aggregation through different levels of hierarchy on the atomic and molecular scale with the novel synergistic use of molecular modeling, molecular dynamics simulation, isothermal titration calorimetry, and other characterization techniques. We are also the fist to elucidate atomic interactions, size, shape, stoichiometry, and association dynamics for polyplexes containing siRNA vs pDNA. Our investigation highlights differences in the hierarchical mechanism of formation of related polycation鈥搒iRNA and polycation鈥損DNA complexes. The results of fluorescence quenching assays indicated a biphasic behavior of siRNA binding with polycations where molecular reorganization of the siRNA within the polycations occurred at lower N/P ratios (nitrogen/phosphorus). Our results, for the first time, emphasize a biphasic behavior in siRNA complexation and the importance of low N/P ratios, which allow for excellent siRNA delivery efficiency. Our investigation highlights the formulation of siRNA complexes from a thermodynamic point of view and opens new perspectives to advance the rational design of new siRNA delivery systems.